Dynamics of dendritic cell‐derived vesicles: high‐resolution flow cytometric analysis of extracellular vesicle quantity and quality

Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society

关键词: exosomes;    microvesicles;    immune regulation;    single vesicle analysis;    cognate interaction;    MHC class II;   

DOI  :  10.1189/jlb.0911480

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

Nano‐sizedmembranevesiclesaresecretedbymanycelltypes.Thesevesiclescanserveascarriersofcellularinformation.DC‐derivedvesiclescanbetargetedtootherimmunecellsandmodifytheirfunction.AccurateanalysisofquantitativeandqualitativechangesinEVproductionbyDCupondifferentactivationstimuliisneededtofurtherrevealtheimmuneregulatorypropertiesofDC‐derivedEVs.However,methodsforreliablequantificationofindividualEVsandforanalysisoftheheterogeneityofEVpopulationsarelimited.Withourrecentlydevelopedhigh‐resolutionflowcytometry‐basedmethod,wecanperformahigh‐throughput,multiparameter,andquantitativeanalysisofindividualEVs.Withtheuseofthisnoveltechnique,weshowthatdespitepreviousassumptions,stimulationwithbacterialLPSincreasesEVreleasebyDC.Furthermore,wedemonstrateheterogeneityinDC‐derivedEVsregardingtheirbuoyantdensityandMHCclassIIcontent.Finally,weshowthatcognateinteractionbetweenLPS‐stimulatedDCandCD4+TcellsaffectsboththequantityandqualityofLPSDC‐derivedEVspresentintheculturesupernatant.Thesedataindicatethatflowcytometry‐basedanalysisofindividualEVsisavaluable,noveltooltostudythedynamicsofEVsecretionandcomposition,offeringgreatopportunitiestounveilthefunctionofimmunecell‐derivedEVs...

【 授权许可】

CC BY